Molecular imaging of bone marrow mononuclear cell survival and homing in murine peripheral artery disease.

OBJECTIVES This study aims to provide insight into cellular kinetics using molecular imaging after different transplantation methods of bone marrow-derived mononuclear cells (MNCs) in a mouse model of peripheral artery disease (PAD). BACKGROUND MNC therapy is a promising treatment for PAD. Although clinical translation has already been established, there is a lack of knowledge about cell behavior after transplantation and about the mechanism whereby MNC therapy might ameliorate complaints of PAD. METHODS MNCs were isolated from F6 transgenic mice (FVB background) that express firefly luciferase (Fluc) and green fluorescence protein (GFP). Male FVB and C57Bl6 mice (n = 50) underwent femoral artery ligation and were randomized into 4 groups receiving the following: 1) single intramuscular (IM) injection of 2 × 10(6) MNCs; 2) 4 weekly IM injections of 5 × 10(5) MNCs; 3) 2 × 10(6) MNCs intravenously; and 4) phosphate-buffered saline as control. Cells were characterized by flow cytometry and in vitro bioluminescence imaging (BLI). Cell survival, proliferation, and migration were monitored by in vivo BLI, which was validated by ex vivo BLI, post-mortem immunohistochemistry, and flow cytometry. Paw perfusion and neovascularization was measured with laser Doppler perfusion imaging (LDPI) and histology, respectively. RESULTS In vivo BLI revealed near-complete donor cell death 4 weeks after IM transplantation. After intravenous transplantation, BLI revealed that cells migrated to the injured area in the limb, as well as to the liver, spleen, and bone marrow. Ex vivo BLI showed presence of MNCs in the scar tissue and adductor muscle. However, no significant effects on neovascularization were observed, as monitored by LDPI and histology. CONCLUSIONS This is one of the first studies to assess kinetics of transplanted MNCs in PAD using in vivo molecular imaging. MNC survival is short-lived, MNCs do not preferentially home to injured areas, and MNCs do not significantly stimulate perfusion in this particular model.

[1]  H. Vogel,et al.  Timing of Bone Marrow Cell Delivery Has Minimal Effects on Cell Viability and Cardiac Recovery After Myocardial Infarction , 2010, Circulation Cardiovascular Imaging.

[2]  K. Shimada,et al.  Therapeutic angiogenesis for patients with limb ischaemia by autologous transplantation of bone-marrow cells: a pilot study and a randomised controlled trial , 2002, The Lancet.

[3]  Stefanie Dimmeler,et al.  Relevance of Monocytic Features for Neovascularization Capacity of Circulating Endothelial Progenitor Cells , 2003, Circulation.

[4]  J. Willerson,et al.  Intra-Arterial Transplantation of Adult Bone Marrow Cells Restores Blood Flow and Regenerates Skeletal Muscle in Ischemic Limbs , 2009, Vascular and endovascular surgery.

[5]  Rutger-Jan Swijnenburg,et al.  Comparison of Different Adult Stem Cell Types for Treatment of Myocardial Ischemia , 2008, Circulation.

[6]  K. Furie,et al.  Heart disease and stroke statistics--2007 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. , 2008, Circulation.

[7]  David M. Bodine,et al.  Bone marrow cells regenerate infarcted myocardium , 2001, Nature.

[8]  L. Norgren,et al.  Inter-Society Consensus for the Management of Peripheral Arterial Disease (TASC II). , 2007, Journal of vascular surgery.

[9]  Y. Natkunam,et al.  Embryonic Stem Cell–Derived Endothelial Cells Engraft Into the Ischemic Hindlimb and Restore Perfusion , 2009, Arteriosclerosis, thrombosis, and vascular biology.

[10]  A. Nagy,et al.  Donor hematopoietic cells from transgenic mice that express GFP are immunogenic in immunocompetent recipients , 2005, Hematology.

[11]  S. Gambhir,et al.  Molecular Imaging of Cardiac Cell Transplantation in Living Animals Using Optical Bioluminescence and Positron Emission Tomography , 2003, Circulation.

[12]  R. Robbins,et al.  Stem cell transplantation: the lung barrier. , 2007, Transplantation proceedings.

[13]  P. Quax,et al.  Vascular growth in ischemic limbs: a review of mechanisms and possible therapeutic stimulation. , 2008, Annals of vascular surgery.

[14]  A. Luttun,et al.  Multipotent adult progenitor cells sustain function of ischemic limbs in mice. , 2008, The Journal of clinical investigation.

[15]  I. Weissman,et al.  Haematopoietic stem cells adopt mature haematopoietic fates in ischaemic myocardium , 2004, Nature.

[16]  P. Christian,et al.  Improved detection of upper abdominal abscesses by combination of 99mTc sulfur colloid and 111In leukocyte scanning. , 1985, AJR. American journal of roentgenology.

[17]  P. Nguyen,et al.  Imaging: guiding the clinical translation of cardiac stem cell therapy. , 2011, Circulation research.

[18]  P. Quax,et al.  Expression of Vascular Endothelial Growth Factor, Stromal Cell-Derived Factor-1, and CXCR4 in Human Limb Muscle With Acute and Chronic Ischemia , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[19]  M. Kay,et al.  Double Knockdown of Prolyl Hydroxylase and Factor-Inhibiting Hypoxia-Inducible Factor With Nonviral Minicircle Gene Therapy Enhances Stem Cell Mobilization and Angiogenesis After Myocardial Infarction , 2011, Circulation.

[20]  M. Gurish,et al.  Pathways of murine mast cell development and trafficking: tracking the roots and routes of the mast cell , 2007, Immunological reviews.

[21]  C. Ye,et al.  Combination of stromal-derived factor-1alpha and vascular endothelial growth factor gene-modified endothelial progenitor cells is more effective for ischemic neovascularization. , 2009, Journal of vascular surgery.

[22]  Jacques Galipeau,et al.  Therapeutic angiogenesis using autologous bone marrow stromal cells: improved blood flow in a chronic limb ischemia model. , 2003, The Annals of thoracic surgery.

[23]  Eric J Topol,et al.  Critical issues in peripheral arterial disease detection and management: a call to action. , 2003, Archives of internal medicine.

[24]  Irving L. Weissman,et al.  Shifting foci of hematopoiesis during reconstitution from single stem cells , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[25]  J. Hamming,et al.  Variations in surgical procedures for hind limb ischaemia mouse models result in differences in collateral formation. , 2010, European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery.